Search

Ruta angustifolia is one of the species of Ruta genus that is widely used in Asia as traditional natural medicine, but scientific research related to this plant is still limited. Several studies have stated that this plant has an antimicrobial effect, but no studies have discussed the antibiofilm effect of bioactive compounds isolated from R. angustifolia. This study is the first to address this issue. This study aims to see whether the bioactive compounds isolated from R. angustifolia can inhibit the formation of biofilms on oral microbes that play the most important role in the construction of dental caries, namely Candida albicans and Streptococcus mutans. Isolation and identification of bioactive compounds is achieved through Thin Layer Chromatography, Liquid Chromatography with tandem mass spectrometry, and proton nuclear magnetic resonance, respectively. Crystal Violet (CV) method is used to see the total biomass of biofilm, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method is used for the viability test. The morphological visualization of C. albicans has also been carried out using a light microscope and a scanning electron microscope. Three compounds that have been isolated are kokusaginine, chalepin, and lindelofine. The CV and MTT data were compared with the control data using an analysis of variance test followed by the Tukey honestly significant difference test, while the morphological data is qualitatively analyzed. From the results of the biofilm test, it was concluded that the three compounds could significantly inhibit the formation of biofilms produced by C. albicans and S. mutans in mixed species cultures.

Context. Proteins in the saliva are one of the defense mechanism factors that can protect the oral cavity from disease. However, smoking might affect the properties of saliva. Aim To determine the differences in salivary protein profiles and total concentrations in smokers and non-smokers and their correlation with dental caries severity as indicated by the Decayed, Missing, Filled-Teeth (DMF-T) scores. Methods and material This cross-sectional study included 25 smokers and 25 non-smokers. The DMF-T scores were recorded. The total salivary protein was measured by the Bradford method, and the profile proteins were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results The average of salivary protein concentration in smokers was lower than that in non-smokers (551.486 µg/mL versus 765.361 µg/mL), but the difference was not statistically significant (P > 0.05). Further correlation analyses showed a negative correlation between the concentration of proteins based on the extent of smoking. A weak negative correlation was found between protein concentration and DMF-T scores (r = −0.239). Dominant salivary protein bands of 11.6 kDa and 54.5 kDa were found in smokers and 27 kDa, 60 kDa, and 94.5 kDa were found in non-smokers. Conclusion Different protein bands appeared in smokers and non-smokers. There was a weak correlation between protein concentration, DMF-T scores, and the extent of smoking.

Background: Gingiva acts as a barrier to prevent further invasion of pathogens in periodontitis. The gingival structure consists of epithelial tissue and connective tissue. As the aging process continues, there are several changes in the periodontium. Previous studies have tried to investigate the complex interaction between the host immune system and bacteria by using animal models, especially rodents. Objective: The aim of the study was to evaluate the effectiveness of collecting gingival tissue from the palate and retromolar pad. Materials and Methods: The aging experimental model had two age categories of male rodents of 18 and 58 weeks. Tissue was collected from the mandible retromolar pad and palate with full-thickness excision. Tissues were transferred to a complete medium at 4°C. Gingival tissue was cultured in a 37°C culture incubator at 5% CO2. Tissue proliferation was observed on the first, third, and fifth days using the hemocytometer. The cell metabolism rate between the two age categories was checked using the MTT Assay. Two-way ANOVA was used for statistical analysis. Results: Gingival tissues obtained from the experimental models of two age categories were alive, and proliferation was observed. The old rodent group showed no significant result in terms of cell morphology on the first vs. third day (p>0.05), but significant results were found on the first vs. fifth day and third day vs. the fifth day (p Conclusion: Rodent gingival tissue collection from the retromolar pad and palate was found suitable for tissue culture in the aging experimental study.

Background: The available evidence suggests that inflammatory responses, in both systemic and oral tissue, contribute to the pathology of COVID-19 disease. Hence, studies of inflammation biomarkers in oral fluids, such as saliva, might be useful to better specify COVID-19 features. Methods: In the current study, we performed quantitative real-time PCR to measure salivary levels of C-reactive protein (CRP) and interleukin-6 (IL-6) in saliva obtained from patients diagnosed with mild COVID-19, in a diabetic group (DG; n = 10) and a non-diabetic group (NDG; n = 13). All participants were diagnosed with periodontitis, while six participants with periodontitis but not diagnosed with COVID-19 were included as controls. Results: We found increases in salivary total protein levels in both the DG and NDG compared to control patients. In both groups, salivary CRP and IL-6 levels were comparable. Additionally, the levels of salivary CRP were significantly correlated with total proteins, in which a strong and moderate positive correlation was found between DG and NDG, respectively. A linear positive correlation was also noted in the relationship between salivary IL-6 level and total proteins, but the correlation was not significant. Interestingly, the association between salivary CRP and IL-6 levels was positive. However, a moderately significant correlation was only found in COVID-19 patients with diabetes, through which the association was validated by a receiver operating curve. Conclusions: These finding suggest that salivary CRP and IL-6 are particularly relevant as potential non-invasive biomarker for predicting diabetes risk in mild cases of COVID-19 accompanied with periodontitis.

Purpose: The purpose of this study was to determine the quantity of Streptococcus mutans serotype c and Candida albicans antigens from dental plaque and their relationship with the Oral Hygiene Index Score (OHI-S) and Decayed, Missing, and Filled Teeth (DMFT) in Early Childhood Caries (ECC) and Severe Early Childhood Caries (S-ECC). Materials and Methods: This was a cross-sectional study, including 5 ECC and 32 S-ECC subjects. ECC was classified as a DMFT score < 4, and S-ECC was classified as a DMFT score ≥ 4 at 71 months of age. S. mutans serotype c and C. albicans antigens from dental plaque of ECC and S-ECC patients were measured using enzyme-linked immunosorbent assays. The optical density ​​of S. mutans serotype c and C. albicans antigens was determined at 450 nm. Correlations between S. mutans serotype c and C. albicans antigens and OHI-S in ECC and S-ECC were analyzed. Results: There were no significant differences in S. mutans serotype c and C. albicans antigens between ECC and S-ECC patients or between patients with moderate and good OHI-S scores. There was a negative correlation between S. mutans serotype c and C. albicans antigens in ECC patients (r = -0,62; p < 0.05). Conclusion: The increase in C. albicans suppresses S. mutans serotype c growth in ECC but not in S-ECC. The findings of this study could be used to discover the molecular mechanisms involved in bacterial-fungal relationships and the contribution of new technologies to reduce ECC.

ObjectiveChronic periodontitis has been proposed to be linked to coronavirus disease (COVID-19) on the basis of its inflammation mechanism. We aimed to evaluate this association by investigating the expression of Angiotensin Converting Enzyme-2 (ACE2) in periodontal compartments, which contain dysbiosis-associated pathogenic bacteria, and how it can be directly or indirectly involved in exacerbating inflammation in periodontal tissue.Material and MethodsThis observational clinical study included 23 adult hospitalized patients admitted to Universitas Indonesia Hospital with PCR-confirmed COVID-19, while 6 non-COVID-19 participants come to periodontal clinic were included as control. Using real time-PCR (qPCR) and gingival crevicular fluids (GCF) samples from COVID-19 patients with and without diabetes and periodontitis, we assessed the mRNA expression of angiotensin-converting enzyme 2 (ACE2), IL-6, IL-8, complement C3, and LL-37 as well as the relative proportion of Porphyromonas gingivalis, Fusobacterium nucleatum, and Veillonella parvula to represent the dysbiosis condition in periodontal microenvironment. All analyses were performed to determine their relationship.ResultsACE2 mRNA expression was detected in the GCF of periodontitis-COVID-19 patients with and without diabetes. However, only periodontitis-COVID-19 patients with diabetes showed a positive relationship between ACE2 expression and inflammatory conditions in the periodontal microenvironment. In addition, the interplay between pro-inflammatory cytokine (IL-6) and complement C3 could be used as a predictor of the severity of periodontal inflammation in COVID-19 patients with diabetes.ConclusionThe study data show that the SARS-CoV-2 entry gene is expressed in the GCF of patients with COVID-19, and its expression correlates with inflammatory markers.

Background/Aims: Nuclear factor kappa beta (NF-κB) is a member of the transcription factor family, and it plays a key role in coordinating the expression of genes in many chronic inflammatory diseases. The main etiology of endodontic treatment failure is caused by bacteria in root canal, including Enterococcus faecalis. The aim of this study to determine NF-κβ and osteocalcin expressions during the periapical tissue damage due to induction of E. faecalis. Materials and Methods: Fifty-four male rats were randomly divided into two main groups, each of which had three subgroups. In Group C (control), every tooth was induced only by sterile brain–heart infusion broth (BHIb). Group C had three subgroups, namely Group C3 with 3 days inductionprocess, Group C10 with 10 days induction process, and Group C21 with 21 days induction process. In Group T, every tooth was induced by 10 μl of BHI-b E. faecalis ATCC212 (106 CFU). Similarly, Group T also had three subgroups, namely Group T3 with 3 days induction process, Group T10 with 10 days induction process, and Group T21 with 21 days induction process. The animals were sacrificed based on their group schedule and then prepared for histological and immunohistochemical examinations of periapical tissue. Afterward, the expressions of NF-κβ and osteocalcin were calculated on the light microscope. Results: The results revealed that the number of cells expressed of NF-κβ and osteocalcin increased significantly in all sub-groups of Group B induced by E. faecalis compared to the control group. Conclusion: It can be concluded that the number of cells expressed NF-κβ and osteocalcin increase during the periapical tissue damage induced by E. faecalis.

Objective: Halitosis may be caused by several factors, including various types of food, periodontal diseases, layer of tongue bacteria, and systemicdisorders such as diabetes mellitus (DM), which is a chronic disease that affects the health of periodontal tissue. The present study aimed to assessthe association between the quantity of Tannerella forsythia bacteria and the levels of methyl mercaptan and hydrogen sulfide in periodontitis patientswith type 2 DM (T2DM).Methods: Gingival crevicular fluids (GCF) were collected from 20 patients who were divided into those with periodontitis and who were normoglycemic(n=8); those with periodontitis and T2DM (n=8); and healthy controls (n=4). The patients underwent intraoral periodontal tissue examination,including pocket depth, attachment loss, plaque index, calculus index, and papilla bleeding index. The quantity of T. forsythia bacteria was evaluatedusing quantitative real-time reverse transcription–polymerase chain reaction. The relationship between the number of T. forsythia bacteria and thelevels of methyl mercaptan and hydrogen sulfide in the patients was analyzed by Spearman’s correlative tests.Results: There is a weak and non-significant correlation (p>0.05) between the levels of methyl mercaptan and hydrogen sulfide and the quantity ofT. forsythia in the GCF and tongue coating of periodontitis patients with halitosis regardless of the presence of T2DM.Conclusion: This study suggests no significant relationship between the levels of methyl mercaptan and hydrogen sulfide and the quantity ofT. forsythia in periodontitis patients with halitosis and DM.

Objective The primary purpose of this study was to evaluate and compare the microbial loads and pathogenicity traits of oral Candida albicans in denture-wearing (DW; n = 15) and nondenture-wearing (NDW; n = 15) elderly persons. Materials and Methods The fungal counts of the saliva, tongue dorsa, and prosthesis-fitting surfaces of the participants were assessed using real-time polymerase chain reaction to compare the quantity and expression of selected C. albicans biofilm-associated genes (ALS3, HWP1, and YWP1). Statistical Analysis The obtained data were analyzed by one-way analysis of variance, followed by Bartlett’s test. When appropriate, the Student’s t-test was also used; a value of p < 0.05 was considered statistically significant. Results In both groups, the count of C. albicans was found to be significantly higher in saliva than in other oral samples. The expression of the hypha-specific genes (ALS3 and HWP1) in the tongue dorsa was higher in the DW group (p < 0.05), whereas the transcription level of the yeast-specific gene (YWP1) was significantly higher in the NDW group. Conclusion Both tongue dorsa and dentures appear to be sharing factors that are important for C. albicans biofilm growth in abiotic and biotic oral surfaces of the elderly.

Melaleuca cajuputi subsp. cajuputi is one of the Australian Melaleuca species commonly found in Pulau Buru (Maluku, Indonesia). Its oil, the M. cajuputi essential oil (MCEO), has been utilized as the main flavor of the Indonesian functional food, Cajuputs Candy. However, the availability of MCEO is becoming limited. On the other hand, Indonesia has many other potential MCEO sources which can be developed as flavor ingredient. Thus, it is noteworthy to explore these new MCEO sources by studying their sensory characteristics and metabolite profiles. This study was conducted to identify potential metabolites that are correlated to sensory attributes of MCEO by using the metabolomics approach. The metabolite profiles of thirteen MCEOs from different origins were analyzed by gas chromatography-mass spectrometry while sensory analyses on Cajuputs Candy were conducted by difference-from-control and rate-all-that-apply tests. Sixty metabolites from the MCEO were annotated that includes 1,8-cineole, α-terpineol, caryophyllene, α-pinene, and γ-terpinene. Sensory analysis revealed cooling aftertaste and sweet taste as favorable attributes. Further analysis using Orthogonal Partial Least Square indicated that 1,8-cineole and γ-terpinene were correlated with cooling aftertaste, while 1,8-cineole and caryophyllene were also correlated with sweet taste. In contrast, linalool and nerolidol were associated with the feature of the most characteristic manufacturer's products which have unfavorable attributes such as floral, iodophor-like, metallic, and soapy attributes. The identification of these metabolites will be useful for the selection of MCEOs that can potentially be used as flavor.

Streptococcus mutans were competing Streptococcus sanguinis in biofilm formation. As pioneer colonizer, S. sanguinis were able to control S. mutans growth. This study was aimed to explore the ability of sucrose and non-sucrose cajuputs candies (SCC and NSCC) in maintaining the antagonistic relationship between the indigenous oral flora when they grew as dual-species biofilms (S. sanguinis and S. mutans). The flavored candies (SCC and NSCC) contained cajuput and peppermint oils as the flavor which the volatile compounds had been identified. The unflavored candies were made similar to the flavored candy but excluding the flavor. The flavored candies, unflavored candies, and the control were exposed in vitro to the biofilms. The biofilms were examined for biofilm inhibition capacity, DNA amount, and the expression level of spxB mRNA. The biofilm inhibition by flavored candies were higher than the unflavored ones and were significantly different compared to the control. The flavored candies managed to decrease the total DNA amount in the biofilm, but unflavored samples did not. The qPCR assays showed that the exposure of candies did not alter the proportion of S. sanguinis DNA to S. mutans DNA in the biofilms. Meanwhile, spxB mRNA expression indicated the ability of S.sanguinis to control S. mutans growth.

Melaleuca cajuputi essential oil (MCEO) from Pulau Buru (PBR), Indonesia, has been used as a functional flavor on Cajuputs Candy for years. Our recent study has explored thirteen other alternative MCEO sources to be developed as food flavor. However, not all of the MCEO had similarities to PBR MCEO both in their sensory and volatile profiles. This study aimed to identify the aroma-active compounds which would affect the overall aroma perception of the most- and the least-liked MCEO as a flavor ingredient based on the nasal impact frequency (NIF) method. Initial screening was performed to evaluate the overall liking of thirteen MCEO on Cajuputs Candy through a hedonic test, conducted by seventy-eight untrained panelists. The sample obtained from Mojokerto (MOJ) was the most-liked alternatives MCEO in a similar degree of liking to the currently used PBR, while Belu (BEL) was the least-liked. Further analysis using Gas chromatography–Mass spectrometry/Olfactometry (GC-MS/O) was carried out on the MCEO from PBR, MOJ, as well as BEL. The response of nine panelists showed that the overall aroma perception of PBR was contributed by 1,8 cineole (eucalyptus-like, mint, fresh), α-pinene (pine, green, fresh), and ylangene (spicy, fresh, woody) as the main aroma-active compounds. In addition to α-pinene and 1,8 cineole, the unique aroma of MOJ was dominantly contributed by caryophyllene, possessing a woody, sweet, and spicy aroma. The strong floral odor of linalool and nerolidol, the aroma-active compounds of BEL, generated distinct sensory characteristics in comparison to the reference, PBR.

Ti-6Al-4V ELI is one of the most commonly used dental implant restore function. The solution treatment temperature variation can significantly increase the strength, but it is not yet known the effect of these temperature variations on the alloy's biocompatibility properties. Twelve female Sprague Dawley rats were divided into six groups as follows: the treated group, the control group, and the defect group without implant material. In the treated group, the femur bone defect was implanted with as-cast Ti-6Al-4V ELI, 850°C, 950°C, and 1050°C heat-treated Ti-6Al-4V ELI implant material. The rats were euthanized after 30 days postimplantation and evaluated histologically. The results show that the histological scoring of the specimen for femur defect without implant material is 2 (fibrous union and fibrocartilaginous), score with implant as-cast is 2.5, the sample with 850°C heat treatment material is 2.5, 950°C is 2.5, and the temperature at 1050°C is 2.5. The score of 2.5 is between score 2 and score 3: hemorrhage, fibrous union, fibrocartilaginous microhemorrhage, and mineralized cartilage union. In conclusion, there is no effect of different heat treatment temperatures for Ti-6Al-4V ELI implant material in rat bone regeneration's maturation level.

Modification of Ti-6Al-4V through electrochemical anodization method has been investigated on the purpose of generating TiO2 nanotube arrays (TiNTAs) on the surface of Ti-6Al-4V films. The as-anodized samples were calcined in an atmospheric furnace at various temperatures, in the range of 500-800 ºC. The evaluation of biofilm inhibition was performed by an in vitro method with Streptococcus mutans as a bacterium model. FE-SEM imaging confirmed the successful formation of TiO2 nanotube arrays while XRD results implied a phase transformation from anatase to rutile when the calcination temperature was around 600-650 ºC with average crystallite size of 18 nm. Calcination temperature is one of determining factors in the adjustment of crystallinity and morphology of TiO2, which in turn affects its capability to suppress biofilm formation. This study revealed that the best sample for biofilm inhibition was calcined at 600 ºC with a crystallite phase of mostly anatase. This sample managed to improve antibacterial activity of up to five times as compared to the unmodified Ti-6Al-4V. The output of this study is expected to give some insight on a promising alternative for preventing the formation of harmful biofilm on dental implants.

Increasing evidence has shown an association between periodontitis and cognitive impairment. Subgingival microbiota play a great role in periodontitis pathogenesis. However, the correlation between the subgingival microbiome and cognitive impairment remains unclear. This study aimed to evaluate the red and orange complex subgingival microbiome of cognitively impaired and cognitively normal elderly Indonesian subjects with periodontitis. Twenty-eight elderly subjects diagnosed with periodontitis underwent two cognitive examinations using the Hopkins Verbal Learning Test and the Mini-Mental State Examination. Gingival crevicular fluid taken from the periodontal pocket, at a depth between 5 and 7 mm, using a paper point was used as the subgingival samples. The subgingival microbiome in the cognitive impairment group (n = 14) and cognitively normal group (n = 14) was compared using the 16S rRNA Metagenomic iSeq™ 100 Sequencing System. There was β-diversity in the subgingival microbiota between the cognitively impaired and cognitively normal subjects. The metagenomic analysis showed a higher abundance of Porphyromonas and Treponema bacteria in the cognitive impairment group than in the normal cognitive group (p < 0.05). The abundance of Porphyromonas gingivalis and Treponema denticola was higher in the cognitively impaired elderly subjects. The role of P. gingivalis and T. denticola in the pathogenesis of cognitive impairment needs further investigation.

Objective: The studies on the influence of geographical and socio-economic factors on the oral microbiome remain underrepresented. The Indonesia basic health research (RISKESDAS) 2018, showed an increasing trend in non-communicable diseases compared with the previous report in 2013. The prevalence of diabetes, heart disease, hypertension, and obesity are reported to be higher in urban areas than in rural areas. Interestingly, non-communicable diseases were found to be more prevalent in women than men. This pilot study aimed to examine the oral health and oral microbiome derived from tongue samples of healthy Indonesian women from urban and rural areas.Methods: Twenty women aged 21–47 years old from West Jakarta, residents of DKI Jakarta (n = 10) as representative of the urban area, and residents of Ende, Nangapanda, East Nusa Tenggara (n = 10) as representative of the rural area were recruited for this pilot study. The participants were evaluated by the Simplified Oral Hygiene Index (OHI-S) according to the criteria of Greene and Vermillion and divided into three groups. High-throughput DNA sequencing was performed on an Illumina iSeq 100 platform.Results: The principal component analysis displayed a marked difference in the bacterial community profiles between the urban and rural localities. The presence of manifest was associated with increased diversity and an altered oral bacterial community profile in the urban women. Two bacterial taxa were present at significantly higher levels (adjusted p < 0.01) in the urban oral microflora (Genus Prevotella and Leptotricia) could account for this difference irrespective of the individual oral hygiene status. The linear discriminant analysis effect size (LEfSe) analysis revealed several distinct urban biomarkers. At the species level, Leptotrichia wadei, Prevotella melaninogenica, Prevotella jejuni, and P. histicola, show an excellent discriminatory potential for distinguishing the oral microflora in women between urban and rural areas. Further, using SparCC co-occurrence network analysis, the co-occurrence pattern in the dominant core oral microbiome assembly was observed to be specific to its ecological niche between two populations.Conclusions: This is the first pilot study demonstrating the characterization of the oral microbiome in Indonesian women in urban and rural areas. We found that the oral microbiome in women displays distinct patterns consistent with geographic locality. The specific characterization of the microbiota of Indonesian women is likely linked to geographical specific dietary habits, cultural habits, and socio-economic status or the population studied.

Objective This study aimed to validate the use of Ca-apt-1, an RNA aptamer, that we generated previously as a probe for immunostaining of Candida albicans in rat tongue paraffin-fixed tissue sections Material and Methods The performance of Ca-apt-1 as a detector molecule was compared with that of anti-C. albicans polyclonal antibody (PcAb), which was used as a positive control. Immunostaining images were visualized by light microscopy and were analyzed by using ImageJ software. Results Microscopic results demonstrated that Ca-apt-1 specifically recognized and immunostained C. albicans cells of rat tongue candidiasis, with a specificity comparable to that of PcAb. ImageJ analysis showed that the area (pixels) detected by Ca-apt-1 was wider than that detected by the antibody. This indicates that the binding affinity of Ca-apt-1 toward C. albicans was better than that of PcAb on paraffin-embedded tissues. Conclusion This study demonstrates that Ca-apt-1 can be used as a probe for immunostaining of fixed tissue sections for oral candidiasis diagnosis.

Background: Subgingival niche is one biofilm habitat containing rich microbiota, which plays an active role in maintaining the health of periodontal tissue and determining host response. As such, a study of changing subgingival biofilms is important for understanding the effect of a systemic condition. In this study, we compared the occurrence of six bacteria cohabiting in the subgingival area of periodontitis subjects, with (DP, n = 8) and without (NDP, n = 4) diabetes. Methods: The six genus and species of targeted bacteria were confirmed by 16S rRNA amplicon sequencing on MinION nanopore platform. Descriptive statistic was used to describe the obtained data. Results: We found that the six genus and species of targeted bacteria were detected but in different quantities in either group's periodontal pocket. Our data showed that Tannerella forsythia was the most abundant species in subgingival biofilms of the DP group of the red complex bacteria. In contrast, Aggregatibacter sp., which belongs to the phylum of proteobacteria, was present at a relatively lower level. In contrast, Fusobacterium sp., which belongs to orange complex bacteria, showed relative similarities in subgingival biofilms of both groups tested, while Veillonella sp., were abundant in the DP groups. Conclusions: Our data show that the diversity of classic periodontopathogens increased in the subgingival niche of periodontitis subjects with diabetes. It is the first study in Indonesia to apply MinION-based, full-length 16S rRNA amplicon sequencing in periodontitis patients with and without diabetes.

Background: Subgingival niche is one biofilm habitat containing rich microbiota, which plays an active role in maintaining the health of periodontal tissue and determining host response. As such, a study of changing subgingival biofilms is important for understanding the effect of a systemic condition. In this study, we compared the occurrence of six bacteria cohabiting in the subgingival area of periodontitis subjects, with (DP, n = 8) and without (NDP, n = 4) diabetes. Methods: The six genus and species of targeted bacteria were confirmed by 16S rRNA amplicon sequencing on MinION nanopore platform. Descriptive statistic was used to describe the obtained data. Results: We found that the six genus and species of targeted bacteria were detected but in different quantities in either group's periodontal pocket. Our data showed that Tannerella forsythia was the most abundant species in subgingival biofilms of the DP group of the red complex bacteria. In contrast, Aggregatibacter sp., which belongs to the phylum of proteobacteria, was present at a relatively lower level. In contrast, Fusobacterium sp., which belongs to orange complex bacteria, showed relative similarities in subgingival biofilms of both groups tested, while Veillonella sp., were abundant in the DP groups. Conclusions: Our data show that the diversity of classic periodontopathogens increased in the subgingival niche of periodontitis subjects with diabetes. It is the first study in Indonesia to apply MinION-based, full-length 16S rRNA amplicon sequencing in periodontitis patients with and without diabetes.

Porphyromonas gingivalis has virulence factors such as gingipain and lipopolysaccharide, causing bacteremia to reach the brain and activate neuroinflammatory release cytokines. This study analyzed the effect of the co-culture of neuron cells with P. gingivalis coated with anti-P. gingivalis antibodies against cytokines produced by neuron cells. The gene expressions of the TNF, IL1B, iNOS2 in neurons was evaluated using RT-qPCR. The results showed that P. gingivalis coated with anti-P. gingivalis antibody before co-culture with neuron cells could decrease the gene expression of TNF, IL1B, and iNOS2 of neuron cells.

Objective Chitosan nanoparticle (nanochitosan) has a broad antimicrobial spectrum against diverse pathogenic microorganisms. However, its effect on dental caries-associated microorganisms, such as Streptococcus mutans and Candida albicans is yet to be explored. These microorganisms are known for causing early childhood caries. Therefore, this study was aimed at investigating nanochitosan inhibition capacity against dual-species biofilms of S. mutans and C. albicans. In this study, nanochitosan antimicrobial activity is reported against mono and dual biofilm species of S. mutans and/or C. albicans at 3 and 18 h incubation time. Nanochitosan inhibition capacity was observed through biofilm mass quantity and cell viability. Results The present study successfully synthesized nanochitosan with average diameter of approximately 20–30 nm, and also established dual-species biofilms of S. mutans and C. albicans in vitro. With nanochitosan treatment, the cell viability of both microorganisms significantly decreased with the increasing concentration of nanochitosan. There was no significant decrease in biofilm mass both in the dual and single-species biofilms after 3 h of incubation. However, greater inhibition of biofilm was observed at 18 h incubation.

Objective: The objective of this study was to investigate the effectiveness of immunoglobulin Y (IgY) antibodies as a dental caries vaccine by utilizingIgY specific to the quorum-sensing signaling receptor ComD of Streptococcus mutans combined with chitosan in the form of mouthwash.Methods: The effects of a mouthwash containing IgY anti-ComD S. mutans with and without chitosan on biofilm-forming isolates of S. mutans wereinvestigated. Subjects were assigned to rinsing twice daily for 6 days with 15–20 ml mouthwash solution for 30 s. Biofilm formation of S mutansisolated from the patiens was measured using a crystal violet method to determine the optical density at 490 nm.Results: The results indicated that mouthwash containing IgY anti-ComD S. mutans and chitosan significantly enhanced the biofilm formation ofS. mutans. In mouthwash containing IgY anti-ComD S. mutans without chitosan, a reduction in biofilm formation was observed; however, this was notstatistically significant.Conclusions: The mouthwash combination of IgY anti-ComD S. mutans and chitosan enhanced the biofilm formation ability of S. mutans isolated fromcaries and caries-free subjects. Further, research is needed to determine the appropriate concentrations of IgY anti-ComD S. mutans and chitosanrequired to effectively inhibit dental caries.

Objective: To determine the expression of TLR4 and MMP8 in gingival crevicular fluid [GCF] in patients with periodontitis. Material and Methods: Clinical samples were collected from 23 gingival crevicular fluid of periodontal disease subjects (n = 14) and healthy periodontal subjects (n=9). Measurement of Clinical parameters of probing pocket depth (PPD), bleeding on probing (BOP), and clinical attachment loss (CAL) were included as diagnostic criteria. Pocket Depth (PD) and CAL were defined as present if the PPD was ≥ 4 mm and the CAL ≥ 1 mm. Expression of TLR4 and MMP8 in the gingival crevicular fluid of deep pockets (PD≥ 6mm), shallow pockets (PD 4-5 mm) and healthy periodontal sulcus (0-3 mm) were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Statistical analysis to compare the pocket was using Independent t-test and Mann-Whitney test. Correlation between mRNA expression and clinical parameters was analyzed using Spearman’s correlation test. Results: Expression of TLR4 was higher in shallow pockets compared to the control group, but the difference was not statistically significant (p>0.05). The expression of MMP8 was higher in shallow pockets compared to the control group, but the difference was not statistically significant (p>0.05) either. There is no significant correlation between TLR4 and MMP8 with clinical periodontal parameters. Conclusion: TLR4 and MMP8 mRNA expression levels should not be used as a clinical biomarker in periodontitis diagnostic tools.

Background: Lactobacillus reuteri and Lactobacillus casei have been proposed as probiotic bacteria that promote oral health. Objectives: The present study aimed to evaluate the in vitro effects of L. reuteri and L. casei on the biofilm formation of major oral pathogens, Streptococcus mutans and Porphyromonas gingivalis. Materials and Methods: L. casei strain Shirota and L. reuteri ATCC 55730 were isolated from the commercial products and cultured in de Man, Rogosa, and Sharpe broth. Polymerase chain reaction was used to confirm the identity of the species. S. mutans ATCC 25175 and P. gingivalis ATCC 33277 were cultured in brain–heart infusion broth and used for biofilm formation on 96-well microplate platform. The biofilms were treated with the probiotics and appropriate controls in a time-dependent experiment from 15 min to 24 h. The biofilm biomass was evaluated using crystal violet and safranin. Results: The statistical analysis showed a significant reduction in the S. mutans and P. gingivalis biofilms after treatment with the L. reuteri and L. casei probiotics at all incubation times (P < 0.05). Conclusion: The present study demonstrated the potential antibiofilm activity of L. casei strain Shirota and L. reuteri ATCC 55730 against S. mutans and P. gingivalis biofilms in vitro. The foregoing data have formed a basis for future clinical studies to evaluate the beneficial oral health effect of probiotic Lactobacilli strains.

Objective: To analyze osteopontin mRNA expression levels in subjects with periodontitis prior to (baseline) and 7, 14, and 28 days following scaling and root planing (SRP). Material and Methods: Gingival crevicular fluid was collected as clinical samples from four subjects with periodontitis (pocket depth, 4 - 5 mm) aged 35 - 54 years old as well as from three healthy subjects (controls). The osteopontin mRNA expression levels were measured by quantitative real-time polymerase chain reaction. Spearman’s rank correlation between osteopontin levels in gingival crevicular fluid and the modified gingival index (MGI) was also performed. Results: The Wilcoxon signed-rank test showed no significant difference in osteopontin mRNA expression levels between baseline and 28 days following SRP (p=0.068). The Friedman test showed no significant difference in osteopontin mRNA expression levels between baseline and following SRP (7, 14, or 28 days) (p>0.05). Spearman’s rank correlation showed no significant correlation between osteopontin mRNA expression levels and MGI (r=0.087; p=0.749). Conclusion: Following SRP of periodontal tissue, there was a decreasing trend in osteopontin mRNA expression; however, this finding was not statistically significant. Nevertheless, osteopontin can be used as a biomarker to monitor the healing process; however, further studies are required to clarify our results.

Objective: To evaluate the relationship between the mRNA transcription level of Matrix Metalloproteinase-9 (MMP-9) and the selected clinical periodontal healing at one month of scaling and root planing. Material and Methods: A total of six chronic periodontitis patients and one periodontally healthy subject were recruited. The gingival crevicular fluid was collected from all subjects, and the expression level of MMP-9 mRNA was measured by quantitative real-time PCR. Pocket depth, papilla bleeding index, and clinical attachment loss were measured on day 1 at baseline and day 30. Scaling and root planing was performed on day 1. Data were analyzed using SPSS 22.0 software. Results: In comparison to the control, periodontal clinical parameters in the treatment group were significantly reduced after scaling and root planing. MMP-9 mRNA expression did not show a significant change after the 30th day. A weak correlation was noted between the MMP-9 mRNA transcription level and the changed PBI measurement. Conclusion: Scaling and root planing is clinically effective for chronic periodontitis with a 4-6 mm pocket, whereas the expression of MMP-9 mRNA was not altered. Further studies with a more extended observation period are needed to confirm or reject the present findings.

Pandemi covid-19 memberikan dampak yang sangat besar dalam berbagai aspek kehidupan terutama aspek pendidikan, dimana pembelajaran yang sebelumnya dilakukan secara konvensional harus dialihkan sepenuhnya pada pembelajaran daring (online). Untuk menunjang proses pembelajaran tersebut, dibutuhkan komponen-komponen pembelajaran lainnya seperti penggunaan media pembelajaran. Salah satu media yang digunakan dalam pembelajaran daring (online) dimasa pandemi covid-19 adalah pengunaan teknologi aplikasi google classroom, dengan harapan dapat memudahkan guru dalam mengelola pembelajaran dan menyampaikan materi secara tepat dan akurat, sehingga pembelajaran akan lebih mudah. Namun dalam prosesnya, terkadang penggunaan aplikasi teknologi google classroom tidak berjalan efektif sesuai dengan yang diharapkan. Sehingga perlu adanya kajian terkait bagaimana motivasi minat belajar siswa dalam proses pembelajaran melalui daring (online) dengan menggunakan aplikasi teknologi google classroom selama pandemi covid-19 sebagai bahan evaluasi dan langkah perbaikan dalam proses pembelajaran. Tujuan penelitian ini adalah untuk menganalisa dan mengevaluasi penggunaan teknologi berbasis e-learning yaitu google classroom sebagai media pembelajaran terhadap motivasi minat belajar siswa pada pembelajaran Bahasa Inggris dimasa Pandemi covid-19. Penelitian ini menggunakan metode penelitian deskriptif kualitatif dan pendekatan kuantitatif dengan prosedur pengumpulan data menggunakan teknik observasi, wawancara, angket (kuesioner) dan dokumentasi.

Aim This study aims to analyze the number Mutans Streptococci (MS) and its protein profile from the saliva of early childhood caries (ECC) and caries-free subjects. Methods MS counts were cultured from saliva samples, and the protein profile of MS was determined from ECC and caries-free subjects. The number of colonies were counted, and the protein bands with the molecular weight of 13, 29, 39, 41.3, 74, and 95 kDa were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis method. Results We found that the number of colonies from saliva of ECC patients was higher than those caries-free (22.20 × 106 CFU/ml vs. 19.16 × 106 CFU/ml, p

Veillonella spp., Gram-negative obligate anaerobic cocci bacteria, amounts to 3% in the oral cavity, relies on the fermentation of lactate as a carbon and energy source for growth. The bacteria are considered anti-cariogenic as they metabolize lactic acid into propionic acid which increases oral environment’s pH and reduces demineralization rate of tooth structure. Identification of Veillonella spp. using traditional methods is difficult due to the lack of conventional phenotypic and biochemical tests. Thus, the biomolecular methods are suitable for the specific detection and identification of Veillonella spp. One of the biomolecular methods that can be used is real-time Polymerase Chain Reaction (PCR), which the results can be qualitative and quantitative. This study aimed to identify Veillonella spp. in tongue plaque’s and saliva’s samples using Real -time PCR. The DNA of Veillonella spp. derived from 36 samples, 18 samples of tongue plaque and 18 samples of saliva, were extracted using a freeze-thaw method and then quantified by real-time PCR using forward primer 5’-CCG TGA TGG GAT GGA AAC TGC-3’ and reverse primer 5’-CCT TCG CCA CTG GTG TTC TTC-3’. Veillonella spp. in 18 samples of tongue plaque was 3,06 x 107 CFU/ml and in 18 saliva samples was 1,51 x 105 CFU/ml. It was concluded real-time PCR can detect Veillonella spp. from all tongue plaque’s and saliva’s samples.

Ruta angustifolia is a medicinal plant often used in treating various diseases. Several studies have been carried out to reveal the chemical compounds in this plant, and some are effectively used as antimicrobials. Candida albicans is the most common pathogenic fungus in humans and causes many cases of infection. C. albicans can form biofilms on biotic and abiotic surfaces, either as mono or multispecies, that may increase its pathogenicity. In a state of forming biofilms, microbes are surrounded by an extracellular matrix that protects the microbial community from antimicrobial agents. In this study, we examined the effect of R. angustifolia extract on C. albicans biofilm formation. C. albicans preformed biofilms (24 h) were treated with various concentrations of R. angustifolia extract (100%, 50%, 20%) and incubated at 37°C for 24 h. The characteristics of biofilm formation were assessed with microscopic methods using a light microscope. The results indicated that the effect of R. angustifolia extract could inhibit the C. albicans hypha formation at all concentrations treated.

Porphyromonas gingivalis has virulence factors such as gingipain and lipopolysaccharide, causing bacteremia to reach the brain and activate neuroinflammatory release cytokines. This study analyzed the effect of the co-culture of neuron cells with P. gingivalis coated with anti -P. gingivalis antibodies against cytokines produced by neuron cells. The gene expressions of the TNF, IL1B, iNOS2 in neurons was evaluated using RT-qPCR. The results showed that P. gingivalis coated with anti -P. gingivalis antibody before co-culture with neuron cells could decrease the gene expression of TNF, IL1B, and iNOS2 of neuron cells.

Background: A relationship between oral microbiota and susceptibility to SARS-CoV-2 infection has been extensively studied. However, the relationship between oral commensal flora and expression of angiotensin-converting enzyme 2 (ACE2) remains to be established. In this observational study, we collected saliva from patients with COVID-19 and evaluated the relationship between ACE2 expression and Candida albicans as well as with selected gram-negative bacteria (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Veillonella parvula). We investigated how this may be directly or indirectly involved in oral dysbiosis in patients with COVID-19. Methods: We included 23 hospitalized patients admitted to Universitas Indonesia Hospital with PCR-confirmed COVID-19, with six healthy participants serving as controls. Saliva and tongue surface swabs were collected from patients with diabetes (DG) and without diabetes (NDG) and subject controls. Using quantitative PCR (qPCR) we assessed the mRNA expression of ACE2, the abundance of C. albicans, and the transcription levels of its biofilm-associated genes, agglutinin-like protein 3 (ALS3), hyphal wall protein 1 (HWP1), and yeast-form wall protein 1 (YWP1). We also counted the relative proportion of the three selected gram-negative oral bacteria in saliva. All analyses were performed to determine the relationship between ACE2 expression and C. albicans and gram-negative bacteria. Results: ACE2 mRNA expression was significantly higher in tongue swab samples than in saliva. However, no significant difference was observed between the patient groups. Conversely, DG patients had a significantly higher abundance of C. albicans in saliva compared to NDG patients and control group patients. The correlation and sensitivity/specificity relationship between ACE2 expression and C. albicans or the selected oral bacteria were also observed. Conclusions: The data show that ACE2 expression can be detected in saliva of patients with COVID-19 and its association with C. albicans and gram-negative oral bacteria might contribute toward developing an oral dysbiosis based predictor for prognosis of COVID-19 severity.

Background: The COVID-19 pandemic caused by SARS-CoV-2 has claimed thousands of lives worldwide. To enhance knowledge and awareness of COVID-19, considerable online resources have been made available, including aspects related to the dental profession. The study aim was to examine the knowledge, perception, and attitude of dental professionals in Indonesia toward COVID-19. We conducted a survey via a questionnaire created using Google docs and distributed to 632 members of the Indonesian Dental Association in the context of a webinar hosted by the Indonesian Oral Biology Association on first June, 2020.Materials and Methods: The questionnaire consisted of 17 items pertaining to demographic data, knowledge and virus identification, awareness regarding drugs commonly used in dentistry during pandemic and research opportunities. Participants were asked to complete the questionnaire after the webinar by choosing one answer to each question. For the analysis, participants were divided into three groups according to their professional background i.e., employment at national hospital, private hospital, or academic faculty. Data were analyzed using descriptive statistics and expressed as frequencies and percentages. The chi-square test was used to investigate the association between professional activity and the level of knowledge, perceptions, and attitudes about COVID-19.Results: Sixty percent of the participants correctly identified the pathogenesis of the disease. This knowledge was not associated with their professional affiliation (p = 0.95). Sixty-seven percentage had comprehensive knowledge about virus detection methods. This knowledge was not associated with their affiliation either (p = 0.54). Questions regarding drugs of choice, prevention, and the spread of COVID-19 were correctly answered by 89, 96, and 82% of the participants, respectively. Knowledge of these aspects were significantly associated with the professional affiliation (p < 0.05). All respondents were optimistic regarding research opportunities (p < 0.01). Respondents from academics were more interested in joining COVID-19-related research projects with governmental institutions (p < 0.01).Conclusion: Knowledge and awareness of COVID-19 among Indonesian dentists are reasonably good. However, further improvement would be beneficial to manage patients during this pandemic. As the number of COVID-19 cases continue to rise in Indonesia, it is important that dentists keep abreast of the updated knowledge on this moving field. Dentist knowledge on infection control should be strengthened through continuous educational programs.

This study explored the influence of cell-free spent media prepared from Aggregatibacter actinomycetemcomitans LuxS mutant (Aa-LuxS), its wild type strain (Aa-WT), and the laboratory strain (Aa-Y4), on the interaction between Candida albicans and Streptococcus mutans while growing in co-species biofilm for 48 h. By analyzing the results of crystal violet staining, [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] (MTT) assays, and quantitative real-time polymerase chain reaction (qPCR), we found that the presence of Aa-LuxS in treated biofilms did not affect biofilm development, while added Aa-WT or Aa-Y4 resulted in a significant decrease in both biofilm mass and the number of cells. The inhibitory effect of Aa-WT or Aa-Y4 was not dependent on the protein concentration in the spent media tested (1 and 10%). Gene transcription analyses indicated that Aa-WT/Aa-Y4 exhibits comparable inhibitory effects on the expression of hyphal-associated genes (ALS3 and HWP1), but not on the expression of YWP1, which encodes a yeast form of C. albicans. In contrast, except for gtfD, the expression of S. mutans gtfB/C genes encoding glucosyltransferase was not affected in Aa-WT and Aa-Y4 treated biofilms compared to the levels found in Aa-LuxS treated biofilms. Our results indicate that AI-2-containing spent media derived from Aa can reduce biofilm biomass without significantly inhibiting the survival rate of S. mutans.

Quantitative proteomic workflow based on mass spectrometry (MS) is recently developed by the researchers to screen for biomarkers in periodontal diseases comprising periodontitis. Periodontitis is known for chronic inflammatory disease characterized by progressive destruction of the tooth-supporting apparatus, yet has a lack of clear pathobiology based on a discrepancy between specified categories and diagnostic vagueness. The objective of this review was to outlined the accessible information related to proteomics studies on periodontitis. The Preferred Reporting Items for Systematical Reviews and Meta-Analysis (PRISMA) statement guides to acquaint proteomic analysis on periodontal diseases was applied. Three databases were used in this study, such as Pubmed, ScienceDirect and Biomed Central from 2009 up to November 2019. Proteomics analysis platforms that used in the studies were outlined. Upregulated and downregulated proteins findings data were found, in which could be suitable as candidate biomarkers for this disease., Cell biology; Proteins; Biochemistry; Molecular Biology; Dentistry; proteomics; Periodontitis; Periodontal disease; Biomarker

Background: Salivary nitric oxide plays an important role as an antibacterial agent in the oral cavity. Here, we analyze salivary nitric oxide, Simplified Oral Hygiene Index (OHI-S) scores and the salivary flow rate in smokers and non-smokers which has not been done previously. Methods: A cross sectional study included 25 smokers and 25 non-smokers. Their OHI-S results were categorized as “good,” “medium,” or “bad.” Unstimulated saliva samples were collected, and their nitric oxide concentration was measured using the Griess method. Results: The salivary flow rate in smokers was lower, at 0.30 ml/minute, compared to non-smokers who had a salivary flow rate of 0.33 ml/minute. This was statistically insignificant. There was a significant difference in the concentrations of nitric oxide between smokers and non-smokers (p < 0.05). Smokers had higher concentrations than non-smokers (185.4 µM Vs 114.60 µM). In addition, there was a moderate positive correlation (r = 0.305) between the concentration of salivary nitric oxide level and the OHI-S results. Conclusions: It was concluded that salivary nitric oxide concentration was higher in smokers, and the oral hygiene condition of smokers was poor.

Background: One of the most interesting effects of probiotics is their ability to modulate the immune system through the induction of cytokines and to enhance the host immune response. Aims: The purpose of this study was to evaluate the anti-inflammatory effect of glycerol-supplemented Lactobacillus reuteri on the transcription level of interleukin (IL)-8 and human-beta-defensin (hBD)-2 expressed by epithelial cells after exposure to bacteria. Materials and Methods: The confluent-cultured HaCat cell line (105 cells/mL) was exposed to Streptococcus mutans ATCC-25175 and Porphyromonas gingivalis ATCC-33277 (107 colony-forming units [CFU]/mL) for 24 h and challenged with probiotic L. reuteri ATCC-55730 (107 CFU/mL) supplemented with glycerol. Subsequently, the transcription levels of IL-8 and hBD-2 in HaCat cells were analyzed using reverse-transcription polymerase chain reaction (RT-PCR). In addition, cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. All the obtained data were statistically analyzed using the one-way analysis of variance test, with P < 0.05 set as the level of significance. Results: The MTT assays confirmed no cytotoxic effects of glycerol-supplemented L. reuteri on HaCat cells (viability >90%). mRNA expression of IL-8 and hBD-2 increased after exposure to both bacteria. The presence of glycerol-supplemented L. reuteri significantly reduced the expression of IL-8 and hBD-2 on HaCat cells (P < 0.05). Conclusion: Glycerol-supplemented L. reuteri reduced the expression of IL-8 and hBD-2, and the results may be proof of principle for a probiotic approach to combating inflammation. However, further studies are needed to validate this probiotic effect.

Betle leaf (Piper betle L.) essential oil and catechu nut (Areca catechu L.) extracts have been known to be able to inhibit biofilm formation of S. mutans. This research aimed to characterize the chemical compounds of betle leaf esssential oil, screen the phytochemicals in catechu nut ethanol extract, and assess the inhibitory potential of betle and catechu in chewy candy on biofilm formation by S. mutans. The experiment included preparation of extracts and chemical characterization of the raw materials, formulation of chewy candy, measurement of biofilm inhibition, and sensory evaluation of the candy. In vitro examination for inhibitory potency of betle and catechu chewy candy against biofilm formation S. mutans ATCC 31987 was performed in adhesion phase (4 hours) and active accumulation phase (18 hours). Antibacterial assay was performed in BHI broth media on microplate 96 wells. Crystal violet 0.5% was used to stain the biofilm and Optical Density (OD) was measured at λ 450 nm. The GC-MS analysis detected 32 compounds in the essential oil of betle leaf. The Betle leaf essential oil contained chavicol acetate, isoeugenol, chavibetol acetate, chavicol, and allylcatechol 3.4-diacetate, while catechu nut ethanol extract contained flavonoids and tannins. The components were possibly the inhibitory agents of S. mutans biofilm formation. Chewy candy containing 0.8% betle leaf essential oil and 2.3% catechu nut extract had effective inhibitory potential for S. mutans biofilm formation. Inhibition during adhesion phase was 74.5±0.7%, while that for accumulation phase was 60.8±1.8%. Sensory analysis suggests that the candy was slightly liked by the panelists (5±2).

This study explores the influence of precoating aptamer (Ca-apt1) on C. albicans viability while the fungus was growing in the presence of exposing condensed cigarette smoke (CSC), prepared from clove (CCSC) and non-clove (NCSC) cigarettes, for 48 h. Using qPCR, we found that mRNA expression of adhesion-associated genes (ALS3 and HWP1) was impaired by precoating C. albicans yeast cells with the aptamer. Conversely, the gene transcription was upregulated when aptamer-uncoated yeast was pre-treated with either CSC. In addition, by analysing the result of MTT ([3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay, we found that the presence of added CCSC or NCSC in growth medium for 48 h was significantly enhanced C. albicans biofilm development. However, the presence of precoated aptamer was significantly impaired biofilm development accelerated by the NCSC. The inhibitory effect of the Ca-apt1 was not dependent on the precoated aptamer (1 and 10%). Interestingly, we noted that the enhancer effect of treated CCSC was no longer effective when the yeast had been precoated with 10% aptamer tested. Additionally, light microscopy analysis revealed that precoating aptamer alleviates morphological changes of C. albicans (from yeast to hypha formation) that are enhanced by adding CCSC or NCSC in the growth medium. In conclusion, these results suggest that administration on Ca-ap1 exhibits a significant protective effect on CSC-induced biofilm formation by C. albicans.

Background: Subgingival niche is one biofilm habitat containing rich microbiota, which plays an active role in maintaining the health of periodontal tissue and determining host response. As such, a study of changing subgingival biofilms is important for understanding the effect of a systemic condition. In this study, we compared the occurrence of six bacteria cohabiting in the subgingival area of periodontitis subjects, with (DP, n = 8) and without (NDP, n = 4) diabetes. Methods: The six genus and species of targeted bacteria were confirmed by 16S rRNA amplicon sequencing on MinION nanopore platform. Descriptive statistic was used to describe the obtained data. Results: We found that the six genus and species of targeted bacteria were detected but in different quantities in either group's periodontal pocket. Our data showed that Tannerella forsythia was the most abundant species in subgingival biofilms of the DP group of the red complex bacteria. In contrast, Aggregatibacter sp., which belongs to the phylum of proteobacteria, was present at a relatively lower level. In contrast, Fusobacterium sp., which belongs to orange complex bacteria, showed relative similarities in subgingival biofilms of both groups tested, while Veillonella sp., were abundant in the DP groups. Conclusions: Our data show that the diversity of classic periodontopathogens increased in the subgingival niche of periodontitis subjects with diabetes. It is the first study in Indonesia to apply MinION-based, full-length 16S rRNA amplicon sequencing in periodontitis patients with and without diabetes.

Coronary Heart Disease (CHD) is the major cause of death in most countries in the world. Gram-positive and Gram-negative bacteria have been identified in bacteremia cases and known to have a role in various vascular diseases, including Streptococcus sanguinis which is most frequently isolated from endocarditis patients and often associated with CHD. The purpose of this study was to analyze the number of Streptococcus sanguinis isolated from dental plaque and saliva of subjects with and without CHD. Bacterial colonies isolated from the dental plaque and saliva of 16 subjects without CHD and 8 subjects with CHD were planted in Mitis salivarius agar, and then the DNA was extracted and quantified with a Real-Time PCR technique using 16S rRNA specific primers. The quantification of Real-Time PCR showed that there was a difference in the number of S. sanguinis between the two groups of subjects, but an unpaired T-test showed that the difference was not statistically significant. Furthermore, the number of S. sanguinis from dental plaque in CHD subjects tends to be higher than that of non-CHD subjects whereas the number of S. sanguinis from saliva in non-CHD subjects tends to be higher than that of CHD subjects.

Y Soeroso,1,2 T Akase,3 H Sunarto,1,2 Y Kemal,1 R Salim,1 M Octavia,1 A Viandita,1 J Setiawan,1 BM Bachtiar2,4 1Department of Periodontology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia; 2Oral Sciences Research Center, Universitas Indonesia, Jakarta, Indonesia; 3R&D, Oral Care Company, Sunstar Group, Takatsuki City, Japan; 4Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia Background: The aim of this study was to evaluate the clinical and microbiological effects of local application minocycline HCl 2% gel, used as an adjunct to scaling and root planing (SRP) for treatment of chronic periodontitis (CP). CP is an inflammation of periodontal tissue that is caused mainly by bacterial infection, where periodontal destruction such as loss of attachment and bone destruction occurred. Methods: A total of 81 subjects with moderate to severe periodontitis whose baseline clinical attachment loss (CAL) was ≥4 mm were randomly assigned to receive SRP alone (control group, N=39) or SRP followed by four times of local application of minocycline HCl gel (Periocline) once a week (test group, N=42). Pocket depth, CAL, and papilla bleeding index were examined at baseline, 21 days, 2, 3, and 6 months. Subgingival plaque samples were collected with sterile curettes and were analyzed by real-time polymerase chain reaction for the presence of three periodontal pathogens (Porphyromonas gingivalis [P.g.], Tannerella forsythia [T.f.], and Treponema denticola [T.d.]) at baseline, 2, 3, and 6 months. Results: The number of bacteria was reduced in both groups at 2 months after baseline (SRP treatment). The changes (2–6 months) in T.d. and T.f. counts in the test group were significantly lower than those in the control group. In the control group, a significant regrowth of P.g., T.f., and T.d. was observed from 2 to 6 months and of P.g. and T.f. from 3 to 6 months. On the other hand, in the test group, the number of the three bacteria did not significantly increase during the 6-month period. Conclusion: The results showed that local application of minocycline, used as an adjunct to SRP, was effective for suppressing regrowth of periodontal pathogens, suggesting its risk reduction of recurrent periodontal pathogens in CP. Keywords: chronic periodontitis, minocycline, scaling and root planing, real-time PCR, periodontal pathogen

This paper investigates the ability of biomachined lab-on-a-chip (LoC) to perform drug testing of Amphotericin B to the Candida albicans. The chip is made of polydimethyl siloxane (PDMS). Molds are patterned using CNC milling followed by biomachining. CNC milling process creates channel features on the bottom mold, while biomachining forms rough surface on the channels. After the molds are created, LoC can be manufactured using those molds. Hence, contour surface on LoC’s channels is also realized following the mold surface. Later, Candida albicans is seeded on the LoC’s channels for 24 and 48 hours with the continuous flow of Yeast Nitrogen Base (YNB) Sterile. Then, cell viability is tested using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium (MTT).The result shows that C. albicans could adhere and grow in the LoC channels. Based on this result, drug testing is conducted in the presence and absence of Amphotericin B (Amp B) under two schemes: without (static) and with (dynamic) the continuous flow of YNB Sterile and Amp B. After 48 hour incubation period, C. albicans biofilm of 28.72 % is shown during dynamic scheme, whereas static scheme had C. albicans biofilm of 99.32 % indicating that the dynamic scheme provides a better efficacy compared to the static scheme.

There is a tendency back to nature treatment in Indonesian community because modern/synthetic medicines is expensive and have a various alarming side effects. That is why natural products medicine becomes more popular, aside from mass media promotions. Researches on natural products such as herbal medicine increases along with the desire to get phytofarmaca that can compete with modern medicines. The aims of this literature review are to discuss and provides information on pharmaceutical standardization of natural products toward phytopharmaca for developing Indonesian traditional medicine. Pharmaceutical standardization will strengthen the basic of pharmaceutical science in producing safe, efficacy, and qualify traditional medicine products. The pharmaceutical standardization consists of standardizing simplicial quality, safety, efficacy, and stable formulation of medicines for use in health services.